Reese J A, Byard J L
In Vitro. 1981 Nov;17(11):935-40. doi: 10.1007/BF02618417.
Hepatocytes were isolated from liver biopsies of rats, guinea pigs, rabbits, dogs, and humans. The procedure is based on cannulation of large veins in the cut face of the biopsy, followed by collagenase perfusion. Yields averaged 19 x 10(6) viable hepatocytes/g liver. Viability averaged 84%, as determined by trypan blue dye exclusion. Cultures were prepared from the isolated hepatocytes and were found to be comparable in morphology and N-demethylase activity to hepatocyte cultures prepared by the in situ perfusion of the liver. The development of this method should facilitate comparative studies of the cytotoxicity, genotoxicity, and metabolism of foreign chemicals in primary hepatocyte cultures.
从大鼠、豚鼠、兔子、狗和人类的肝脏活检组织中分离出肝细胞。该方法基于在活检组织切面的大静脉插管，随后进行胶原酶灌注。平均产量为每克肝脏19×10⁶个活肝细胞。通过台盼蓝染料排斥法测定，存活率平均为84%。从分离出的肝细胞制备培养物，发现其在形态和N-脱甲基酶活性方面与通过肝脏原位灌注制备的肝细胞培养物相当。该方法的开发应有助于在原代肝细胞培养物中对外源化学物质的细胞毒性、遗传毒性和代谢进行比较研究。
